Mechanisms of Tolerance To Salinity in Banana: Physiological, Biochemical, and Molecular Aspects

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ISSN 0100-2945 http://dx.doi.org/10.1590/0100-29452017723

MECHANISMS OF TOLERANCE TO SALINITY IN BANANA:


PHYSIOLOGICAL, BIOCHEMICAL,
AND MOLECULAR ASPECTS1
Lilia Willadino2, Terezinha Rangel Camara3, Marta Barbosa Ribeiro4,
Daniel Oliveira Jordão do Amaral5, Flavia Suassuna6, Márcia Vanusa da Silva7

ABSTRACT - In the northeastern region of Brazil, saline soils are constraints to banana production,
becoming necessary to understand the mechanisms of salt tolerance. Two bananas genotypes, Tap Maeo,
tolerant, and Berlin, sensitive, were subjected to treatment with 50 mol m-3 NaCl or without salt. This study
evaluated the effects of salt on the following physiological aspects: leaf area, content and distribution of
Na+, membrane integrity, proton ATPase activity. Besides, a search for differentially expressed genes was
performed using the Differential Display technique. Tap Maeo genotype showed the smallest reduction in
leaf area, smaller accumulation of Na+ and malondialdehyde (MDA), and higher activity of proton ATPase
activity. Two sequences differentially expressed in the tolerant genotype, (Musa 07, Musa 23), shared a
high degree of identity with the amino acid sequences of the genes SOS1 and SOS2, respectively. The clone
Musa 10 was highly similar to amino acid sequence of the ascorbate peroxidase gene, and Musa 26, encodes
the enzyme betaine aldehyde dehydrogenase. These significant biological markers indicate that salinity
tolerance in banana involves at least two simultaneous mechanisms: the activation of the SOS pathway,
increasing the extrusion of Na+, and the activation of antioxidative system, increasing the synthesis of APX
and betaine aldehyde dehydrogenase enzyme.
Index terms: DDRT-PCR, leaf area, Musa, Na+, antioxidative system, SOS pathway.

MECANISMOS DE TOLERÂNCIA À SALINIDADE EM BANANEIRA :


ASPECTOS FISIOLÓGICOS, BIOQUÍMICOS E MOLECULARES
RESUMO - Na região Nordeste do Brasil, solos salinos são restritivos à produção de banana, tornando-
se necessário compreender os mecanismos de tolerância de sal. Dois genótipos de banana, Tap Maeo,
tolerante, e Berlim, sensível, foram submetidos ao tratamento com 50 mol m-3 NaCl ou sem sal. Este estudo
avaliou os efeitos do sal sobre os seguintes aspectos fisiológicos: área foliar, conteúdo e distribuição de Na+,
integridade da membrana, atividade da ATPase. Além disso, uma busca por genes diferencialmente expressos
foi realizada usando a técnica Differential Display. O genótipo Tap Maeo apresentou a menor redução na
área foliar, menor acúmulo de Na+ e malondialdeído (MDA) bem como maior atividade da H+ATPase.
Duas sequências diferencialmente expressas no genótipo tolerante (Musa 07, Musa 23) compartilham alto
grau de identidade com as sequências de aminoácidos dos genes SOS1 e SOS2, respectivamente. O clone
Musa 10 é muito semelhante à sequência de aminoácidos do gene da peroxidase do ascorbato, e o Musa 26
codifica a enzima aldeído betaína desidrogenase. Estes marcadores biológicos significativos indicam que
a tolerância à salinidade em banana envolve pelo menos dois mecanismos simultâneos: a ativação da via
SOS, aumentando a extrusão do Na+, e a ativação do sistema antioxidante, aumentando a síntese de APX e
da enzima aldeído betaína desidrogenase.
Termos para indexação: área foliar, DDRT PCR, Musa, Na+, rota SOS, sistema antioxidativo.
1
(Paper 232-15 ). Received on September 29, 2015. Accepted for publication July 08, 2016.
2
Biologist, PhD. Department of Biology, Federal Rural University of Pernambuco, Avenida Dom Manoel de Medeiros s/n, 52021060,
Recife, Pernambuco, Brasil. E-mail: willadino.lilia@gmail.com (autor para correspondência);
3
Agronomist, PhD. Department of Chemistry, Federal Rural University of Pernambuco, Avenida Dom Manoel de Medeiros s/n,
52021060, Recife, Pernambuco, Brasil. E-mail: teca.camara@gmail.com
4
Forestry Engineer, Master in Chemistry, Department of Chemistry, Federal Rural University of Pernambuco, Avenida Dom Manoel
de Medeiros s/n, 52021060, Recife, Pernambuco, Brasil. E-mail: martaribeiro21@hotmail.com
5
Biologist, PhD. National Institute of the Semi-Arid, Avenida Francisco Lopes de Almeida, s/n, 58429970, Campina Grande, Paraiba,
Brasil. E-mail: danieljornal@yahoo.com.br
6
Biologist, Master in Botany, Department of Biology, Federal Rural University of Pernambuco, Avenida Dom Manoel de Medeiros
s/n, 52021060, Recife, Pernambuco, Brasil. E-mail: flaviasuassuna@yahoo.com.br
7
Agronomist, PhD. Department of Biochemistry, Federal University of Pernambuco, Avenida Prof. Moraes Rego s/n, 50670420,
Recife, Pernambuco, Brasil. E-mail: marciavanusa@yahoo.com.br

Rev. Bras. Frutic., v. 39, n. 2: (e-723) DOI 10.1590/0100-29452017 723 Jaboticabal - SP


2 L. Willadino et al.

physiological relevance and structural simplicity


INTRODUCTION of the root tissues makes them an obvious target
Salinity is one of the most significant abiotic for functional genomic analysis (KUMAR et al.,
stresses in the world, constituting a limiting factor 2015) Several techniques have been developed to
that adversely affects the growth of plants in general identify genes that are differentially expressed and
and the productivity of agricultural crops. In the last that are involved in the mechanism of tolerance
decade, more than 900 million hectares worldwide to abiotic stress in plants (GUPTA; HUANG,
were recorded as having been affected by salinity 2014). Among these techniques, DDRT-PCR, still
(FAO 2008). managing a lower volume of information than the
Plant species, glycophytes or halophytes, that other techniques, has a relatively low cost and is
survive in saline environments exhibit adaptation easy to perform (LIANG; PARDEE, 1995). Several
strategies that involve physiological, biochemical plant-related studies have shown that DDRT-PCR
and molecular mechanisms. Studies have shown can be effectively used to clone genes that are
that plants develop a tolerance to osmotic stress, preferentially expressed in plants by abiotic stress
ionic toxicity stress (FLOWERS et al., 2015), and (UEDA et al., 2002).
oxidative stress (FOYER; SHIGEOKA, 2011). Considering the importance of saline stress
To alleviate Na+ toxicity and re-establish cellular as a limiting factor for banana production and given
homeostasis is necessary the extrusion of Na+ by that the physiological and biochemical specific
the salt overly sensitive pathway (SOS) (PARDO, responses to salinity represent a combination of
2010), and this transport across the membrane is prior molecular events activated by the perception
dependent of proton ATPase activity (VITART of salt, it is necessary to identify and characterize
et al., 2004). The salt SOS pathway functionally the genes involved in the response to salinity. The
consists of the Na+ transporter, SOS1, the protein objective of this study was to isolate, sequence,
kinase SOS2, and the Ca2+ sensor SOS3, which and identify differentially expressed genes in the
constitute a functional model that ensures ionic saline stress-tolerant banana genotype, as well as to
homeostasis in plants that are adapted or tolerant to associate these genes with their physiological and
saline stress (FEKI et al., 2014). Another important biochemical mechanisms.
mechanism for tolerance is the increase in the
activity of antioxidant enzymes, to avoid the effects
of oxidative stress (FOYER; SHIGEOKA 2011). MATERIALS AND METHODS
This is a mechanism that avoid the accumulation
of reactive oxygen species (ROS) that causes
Micropropagated banana plantlets (Musa
damage to nucleic acids, proteins, membrane lipid
sp.) of Tap Maeo and Berlin genotypes, which are
peroxidation and disrupt the redox homeostasis
tolerant to and sensitive to salinity, respectively,
(AZOOZ et al., 2009).
were acclimated for 25 days in a hydroponic system
Tolerance is affected by many different
prior to the experiments. The plants were grown in
genes involved in these various metabolic pathways,
a nutrient solution containing 742.86 mg.L-1 soluble
such as ion extrusion, ion compartmentalization,
fertilizer (3% N, 11% P2O5, 38% K2O, 4% MgO,
synthesis of compatible solutes and scavenging
11% S, 0.025% B, 0.004% Mo, 0.01% Cu-EDTA,
ROS (SHI et al., 2000; SUN et al., 2010; KUMAR
0.025% Zn-EDTA, 0.07% Fe-EDTA, and 0.04%
et al., 2009). Genotypes that differ with regard
Mn-EDTA) and 840.00 mg.L-1 calcium nitrate
to their tolerance for salinity therefore exhibit
(15.5% N and 19.0% Ca). The salt treatment began
qualitative and/or quantitative differences in gene
after the acclimation period by adding 50 mol.m-3
expression. The identification and understanding of
NaCl. The treatment without NaCl was used as the
the molecular control mechanisms involved in the
control.
tolerance to abiotic stresses may result in the use
The plants were analyzed considering the
of molecular tools to produce new more tolerant
time of the expression of the events to be evaluated.
commercial cultivars (GUPTA; HUANG, 2014)
At the beginning of the saline treatment
In many circumstances, it is the sensitivity of the
(time zero) and after 24 and 48 hours, five plants
roots to stress that limits the plant productivity.
in each treatment group were collected. The roots
Despite this direct action of stress on the roots, the
were frozen in liquid nitrogen to extract the total
majority of studies focus primarily on the leaf tissue
RNA and for the DDRT-PCR analysis. Seven days
in experiments involving the exposure of plants
after the initiation of the treatments, five plants in
to saline stress. However, the combination of the
each treatment were collected to evaluate membrane

Rev. Bras. Frutic., v. 39, n. 2: (e-723) DOI 10.1590/0100-29452017 723 Jaboticabal - SP


MECHANISMS OF TOLERANCE TO SALINITY IN BANANA: PHYSIOLOGICAL, ... 3

lipid peroxidation and the H+ATPase activity in the edu/tgi/cgi-bin/tgi/Blast/index.cgi), and the DNA
membrane. After fifteen days of treatment, the leaf Data Bank of Japan (DDBJ; http://www.ddbj.nig.
area was measured, and five plants were collected ac.jp/search/blast). Multiple sequence alignments
to analyze the Na+ and K+ concentrations. were performed using ClustalW software (http://
The experiment was carried out in a www.ebi.ac.uk) (THOMPSON et al., 1994), and
completely randomized design with two levels of a phylogenetic tree was constructed using the
salt (0 and 50 mol.m-3 NaCl), two cultivars (Tap neighbor-joining method with MEGA 3.1 software
Maeo and Berlin) making a total of 4 subplots. Each (KUMAR et al., 2004). The reliability of the tree
subplot contained 10 plants of each genotype, with was assessed by bootstrap analysis with 1500
a total of 40 plants. The experiment was repeated simulations and edited using MEGA 3.1 software.
3 times. The data set was submitted to analysis
of variance (ANOVA) and mean values were
compared using Tukey’s test (P < 0.05).
RESULTS AND DISCUSSION
The leaf area (LA) was determined using
a leaf area meter (Li-Cor, Inc., Lincoln, Nebraska, The salinity reduced the leaf area of both
USA). The analysis of the Na+ and K+ concentrations genotypes by different magnitudes (Table 1). In
was performed using flame photometry. the tolerant genotype, Tap Maeo, the reduction in
The plasma membrane vesicles were isolated leaf area was 21.8%, whereas the reduction in the
by differential centrifugation of the root primordial. sensitive genotype Berlin was 45.2%.
The ATPase activity of the plasma membrane was The leaf area reduction that occurs due to the
determined by measuring the Pi liberated during increase of the levels of salt is a common response
ATP hydrolysis, according to the colorimetric in banana and has been previously described in
method described by Fiske and Subbarrow, with 46 genotypes evaluated by different researchers
the modifications (FAÇANHA; DE MÉIS, 1998). (GOMES et al., 2004; SILVA et al., 2009; FERRAZ,
Vanadate (membrane ATPase inhibitor) and nitrate 2008; WILLADINO et al., 2011); these genotypes
(V-ATPase inhibitor) were used as inhibitors. exhibited reductions in leaf area that varied from
The total RNA was extracted from the roots 16.3 to 69.2%. Decreased leaf area is related to
using TRIzol. For DDRT-PCR, a 1 µg aliquot of constrained CO2 fixation, which reduces the net
total RNA was used as a standard to obtain cDNA assimilation rate and increases the production of
and for the subsequent PCR amplification. The ROS through the Mehler reaction (MILLER et
cDNA synthesis reactions were performed at 50°C al., 2009). The reduction in leaf area is also due to
for 50 minutes. The PCR reactions were performed the deviation of energy required for the activation
using combinations of anchor primers (A1, A2, A3, and maintenance of metabolic activities that are
or A4) with random primers (B1, B2, or B3). The associated with salinity tolerance, such as the
amplification conditions were as follows: 94°C for maintenance of membrane integrity, the synthesis
2 minutes, followed by 40 cycles at 94°C for 30 of organic solutes, ion extrusion, and the regulation
seconds, 42°C for 1 minute, and 72°C for 1 minute, of ion transport and distribution in various organs
as well as a final elongation stage at 72°C for 5 and within cells (MUNNS; TESTER, 2010).
minutes. PCR was performed at least three times for The K+ concentration in the shoots and roots
each primer pair. The PCR-amplified products were varied with the addition of NaCl to the nutrient
separated on 1.5% agarose gels. The genes induced solution. Whereas the sodium concentration
by salt were defined when the expression was 1.5 increased in the roots with increasing salinity but did
fold greater than that of the control, as quantified by not differ between the genotypes. In addition in the
the PCR product. The nucleotide sequences of each shoots, while the Tap Maeo genotype demonstrated
clone were determined using an automatic DNA a small increase in Na+ concentration the Berlin,
sequencer (ABI PRISM 377; Applied Biosystems, sensitive genotype, exhibited an increase of more
Foster City, California, USA), using the Big Dye than 300% (Table 1). This Na+ distribution pattern,
Terminator kit (Applied Biosystems). The BlastX which is characterized by low concentrations of
program (ALTSCHUL et al. 1997) was used to that ion in the shoots is characteristic of salinity
compare the obtained sequences with homologous tolerance, as observed in 30 banana genotypes
sequences in the databases of the National Center exposed to the same conditions (GOMES et al.,
for Biotechnology Information (NCBI; http://www. 2004; GOMES et al., 2005; WILLADINO et al.,
ncbi.nlm.nih.gov/blast), the Institute for Genomic 2011). The maintenance of ionic homeostasis in
Research (TIGR; http://compbio.dfci.harvard. the tolerant genotypes requires the extrusion of Na+

Rev. Bras. Frutic., v. 39, n. 2: (e-723) DOI 10.1590/0100-29452017 723 Jaboticabal - SP


4 L. Willadino et al.

from the cytosol to the vacuole or apoplast through shoot cells of the tolerant genotype Tap Maeo
the Na+/H+ antiporter (SHI et al., 2002). (Table 1), thereby protecting the photosynthetic
The analysis of total RNA revealed apparatus. The cDNA clone Musa 23, which is
fragments corresponding to rRNA 28S and 18S differentially expressed in the Tap Maeo genotype,
with optical quality (A260/230) and density (A260/280) exhibited 73% similarity to the amino acid
ratios of approximately 1.9 and 1.7, suggesting low sequence of the SOS2 gene in Arabidopsis (access
contamination with polysaccharides and proteins, number AF237670.1) (Table 2). Greater expression
respectively, and a high yield of total RNA. of SOS2 is responsible for better salinity tolerance
Twelve primer combinations were used. response in different genotypes under salt stress
The optimal pattern and resolution clarity of the (CHAKRABORTY et al., 2012). The SOS2 gene
amplified products were obtained with the A2B2 encodes a serine/threonine protein kinase with
primer pair 48 hours after stress. either a regulatory or second catalytic domain, with
The transcripts of all of the samples (T0, T1, a regulatory-catalytic interaction within the SOS2
and T2) generated by the same pair of primers were gene. The protein kinase SOS2 interacts with a
applied side-by-side. Twenty-three differentially Ca2+-binding protein similar to calcineurin, SOS3,
expressed amplicons were isolated, cloned, and which responds to an increased Ca2+ concentration
sequenced. The length of the amplicons varied in the cytosol and triggers the interaction between
between 300 and 1900 base pairs (bp). The SOS2 and SOS3 (FEKI et al., 2014).
similarity between the sequences obtained here and The SOS2/SOS3 complex activated by sa-
previously known gene sequences was determined. line stress is responsible for the phosphorylation
Similarity was considered to be present when the and activation of SOS1, the Na+/H+ antiporter,
E values were less than (1e-05), which would which is present in the membrane (MILLER et al.,
therefore be considered significant. 2009). Therefore, the SOS2 and SOS3 proteins are
The tolerant variety, Tap Maeo, have the components of the SOS pathway. The SOS path-
differentially expressed Musa 07 cDNA fragment way ensures the ionic homeostasis of the Tap Maeo
(Table 2) in the roots that have a high homology to genotype through the extrusion of Na+ from the
SOS1 gene that encoding SOS1, the Na+/H+ antiport. cytoplasm to the apoplast or vacuole. The Na+/H+
This cDNA fragment (approximately antiporter is a secondary transporter that depends
500 bp) have an amino acid sequence with 85% on the activity of ATPases the primary transporters.
homology to the Salt Overly Sensitive 1 (SOS1) ATPases are responsible for the electrochemical
gene of the glycophyte Arabidopsis thaliana (access gradient that ensures the energization of the Na+/H+
number, AF256224) (SHI et al., 2000) (Figure 1) antiporter (VITART et al., 2004). The plants of the
and 81% homology to Pisum sativum, Thellungiella Tap Maeo genotype exhibited an increase of more
halophile, and Brassica napus. than 20% in the activity of ATPase and PPase, en-
The salt overly sensitive (SOS) pathway suring sodium extrusion, and the sensitive genotype
functionally consists of the Na+ transporter, SOS1, demonstrated a decrease in the activity of these pro-
the protein kinase SOS2, and the Ca2+ sensor SOS3, ton pumps (Figure 3).These significant biological
which constitute a functional model that ensures markers indicate that activation of the SOS pathway
cellular homeostasis in plants that are adapted or is a fundamental mechanism for saline stress toler-
tolerant to saline stress (FEKI et al., 2014; KUMAR ance of banana.
et al., 2009). The phylogenetic tree of the Na+/H+ The cDNA fragment Musa 26 encodes be-
antiport sequences (SOS1), based on higher plants, taine aldehyde protein dehydrogenase (BADH, EC
animals, fungi, and Escherichia coli, indicated 1.2.1.8), enzyme that catalyzes the oxidation of be-
two distinct “clusters”, as supported by the high taine aldehyde to glycine betaine (GB). GB plays
bootstrap value, i.e., a vacuole cluster and a plasma important role in stress tolerance that include the
membrane cluster, which indicates that Musa 07 is stabilization of complex proteins and membranes
a fragment of the Na+/H+ antiporter gene (Figure 2). in vivo, protection of the transcriptional and trans-
This same antiporter is also involved in the lational machinery, reduction of ROS accumula-
distribution of Na+ between the shoots and the root tion and peroxidation of membrane lipids, and as
(PARDO, 2010) by restricting xylem loading with a molecular chaperone in the refolding of enzymes
the Na+ ion, thereby preventing its transportation (GIRI, 2011; CHEN; MURATA, 2011). The per-
to the shoots (SUN et al., 2010; SHI et al., 2002). oxidation of membrane lipids was monitored indi-
This altered distribution of Na+ prevents toxic rectly through the production of malondialdehyde
concentrations of this cation in the cytoplasm of (MDA), a byproduct of lipid peroxidation. The Tap

Rev. Bras. Frutic., v. 39, n. 2: (e-723) DOI 10.1590/0100-29452017 723 Jaboticabal - SP


MECHANISMS OF TOLERANCE TO SALINITY IN BANANA: PHYSIOLOGICAL, ... 5

Maeo genotype did not exhibit variations in the production of H2O2 and other ROS (O2-, OH-, and
MDA concentration when subjected to the stress, 1
O2) is cytotoxic (FOYER; SHIGEOKA, 2011)
however, in the Berlin genotype, the accumulation and frequently occurs in plants subjected to bi-
of Na+ caused an increase in the concentration of otic or abiotic stress (MITTLER, 2006). Tolerant
MDA (Figure 3). genotypes respond to such stress with an increase
The ability of tolerant varieties to maintain in the activity of antioxidant enzymes, preventing
low concentrations of MDA, as well as MDA ac- and avoiding damage to nucleic acids, proteins and
cumulation in susceptible varieties, indicates that membrane lipid peroxidation, thus maintaining re-
variations in lipid peroxidation of membrane can dox homeostasis (AZOOZ et al., 2009). The differ-
be used to evaluate the effect of salt stress on cell, ences on the biochemistry and molecular markers
once the membrane is the first site to perceive this highlights the importance of the antioxidant system
stress. Another cDNA fragment, the Musa 10 clone, as a mechanism of salt stress tolerance in banana.
was identified and exhibited 71% similarity to the In summary, salinity tolerance in Musa in-
amino acid sequence of the ascorbate peroxidase volves a set of at least two simultaneous mecha-
(APX) gene of Vigna unguiculata (access number nisms. These mechanisms include the activation of
U61379.1). APX is the most important enzyme as- the SOS system, which ensures the extrusion of Na+
sociated with the elimination of H2O2 from the cy- from the cytoplasm and the activation of the anti-
tosol and chloroplasts (BARBOSA et al., 2014), oxidative system, in particular the increase in the
preventing the oxidative damage. The excessive synthesis of the enzyme APX and glycine betaine.

Table 1 - Leaf area (LA), Na+ and K+ concentrations (g.kg-1), and K+/Na+ ratio in shoots (S) and roots
(R) of banana genotypes grown for 15 days with or without 50 mol m-3 NaCl in the nutrient
solution
Treatment LA Na+ K+ K+/Na+
Genotype
(mol m-3 NaCl) (cm2) S R S R S R
0 3.26 a 3.7 c 4.2 b 31.2 a 41.4 a 8.4 9.8
Tap Maeo
50 2.53 b 5.9 b 6.2 a 29.7 a 40.9 a 5.0 6.6
0 2.79 b 3.4 c 3.8 b 32.3 a 40.6 a 9.5 10.7
Berlin
50 1.26 c 16.5 a 6.9 a 30.0 a 39.9 a 1.8 5.8
LA Reduction Na+ Increase Reduction (%)
(%) (%) K+ K+/Na+
Tap Maeo 21.8 59.5 47.2 4.6 0.2 49.5 32.7
Berlin 45.2 385 81.6 7.1 1.7 89.5 45.8
Data indicated by different letters within each column exhibit significant difference at the 5 % probability level according to Tukey’s test

Table 2 - Catalog of cDNA detected by differential display in banana roots*.


PCR Identity of
N° cDNA Primer Access Best homology
Fragment Organism aminoacid
fragment combinations number to database
size (bp) (%)
Musa putative Na+/H+ Arabidopsis
A2B2 500 AF256224.1 85
07 antiporter SOS1 thaliana
Musa cytosolic ascorbate Vigna
A2B2 390 U61379.1 81
10 peroxidase unguiculata
Musa serine/threonine Arabidopsis
A2B2 430 AF237670.1 73
23 protein kinase SOS2 thaliana
betaine aldehyde
Musa Suaeda
A2B2 450 AF359282.1 dehydrogenase 72
26 liaotungensis
(BADH)
* Of the sequences obtained, four exhibited a high degree of similarity to amino acids sequence of genes whose functions are already
known, nine displayed low similarity, and ten sequences were not similar

Rev. Bras. Frutic., v. 39, n. 2: (e-723) DOI 10.1590/0100-29452017 723 Jaboticabal - SP


6 L. Willadino et al.

Figure 1 - Comparison of amino acid sequence between Musa 07 clone of Musa sp. with the SOS1 gene of
A. thaliana (AF256224). Sequences labeled with colons, asterisks, periods, and dashes represent those with
similar amino acids, identical amino acids, different amino acids, and a lack of amino acids, respectively.
The ClustalW program was used to generate the alignment (THOMPSON et al., 1997).

Figure 2 - Musa 07 clone position in the phylogenic tree of Na+/H+ antiporter. Multiple alignments of the sequences
were performed with CLUSTALW, and a phylogenic tree was built using the neighbor-joining method with the MEGA
3.1 program. The accession numbers (in parentheses) and the Na+/H+ antiporter sources are as follows: AgNHX1
(AB038492), Atriplex gmelini; AtNHX1 (AF510074), AtSOS1 (AF256224), and AtSOS1 (AY062746), Arabidopsis
thaliana; NHE2 (AAD41635) and NHE5 (AAC98696), Homo sapiens; OsNHA1 (AY328087), Oryza sativa; TaSOS1
(AY326952), Triticum aestivum; PpSOS1 (AJ564258), Physcomitrella patens; PeNhaD1 (AJ561195), Populus
euphratica; NhaP (BAA31695), Pseudomonas aeruginosa; NHA1 (NP-013239), Saccharomyces cerevisiae; SOD2
(CAA77796), Schizosaccharomyces pombe; NhaA (P13738), Escherichia coli; and PeSOS1 (DQ517530), Populus
euphratica

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MECHANISMS OF TOLERANCE TO SALINITY IN BANANA: PHYSIOLOGICAL, ... 7

Figure 3 - Hydrolytic activity of P-ATPase and PPiase and MDA concentration in roots of banana
genotypes grown for 7 days with or without 50 mol m-3 NaCl in the nutrient solution.
Significant difference between the means of the treatments according to F test (0,05%).

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